雏鸡冷应激前后血清中差异蛋白质组学研究

发布时间：2018-05-15 05:08

本文选题：双向电泳 + 冷应激　；参考：《动物营养学报》2017年05期

【摘要】：本试验旨在比较分析雏鸡冷应激前后血清中蛋白质的表达差异,并对重点差异蛋白质进行鉴定。将30只雏鸡随机分为3组,分别为冷应激组、冷适应组和常温对照组,收集各组雏鸡的血液制备血清后进行双向凝胶电泳(2-DE),以获得血清蛋白质表达的2-DE图谱,对2-DE图谱进行差异分析(利用PDQuest 8.0软件),随后采用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)技术对差异表达蛋白质进行鉴定,并利用蛋白质印迹(Western blot)方法进行验证。结果显示:通过2-DE对常温对照组、冷应激组和冷适应组雏鸡血清进行分析,得到了比较完整的差异蛋白质数据,共找到差异蛋白质点23个。采用MALDI-TOF-MS技术分析其中几个重复性好且较为明显的蛋白质点,成功鉴定出4个差异蛋白质,其中2个为果糖二磷酸醛缩酶C(ALDOC),是参与葡萄糖、能量代谢通路供能的相关蛋白;随后,对差异蛋白质ALDOC进行Western blot验证,所得结果与2-DE的结果相一致。结果表明,雏鸡冷应激前后血清中蛋白质的表达具有明显的差异,这些蛋白质的表达差异可能与冷应激有关。
[Abstract]:The purpose of this experiment was to compare and analyze the difference of serum protein expression before and after cold stress in chicks, and to identify the key differential proteins. Thirty chicks were randomly divided into three groups: cold stress group, cold acclimated group and normothermic control group. The blood samples of each group were collected to prepare serum for 2-DEG to obtain the 2-DE map of serum protein expression. The differentially expressed proteins were identified by PDQuest 8.0 software and then identified by MALDI-TOF-MS, and verified by Western blotting. The results showed that the serum of chicks in normal temperature control group, cold stress group and cold acclimation group were analyzed by 2-DE, and complete differential protein data were obtained, and 23 differential protein spots were found. MALDI-TOF-MS technique was used to analyze several protein spots with good reproducibility, and four differentially expressed proteins were successfully identified, two of which were fructose diphosphate aldolase, which was related to the energy supply of glucose and energy metabolism pathway. The differential protein ALDOC was verified by Western blot, and the result was consistent with that of 2-DE. The results showed that there were significant differences in the expression of serum proteins in chicks before and after cold stress, and the differences in the expression of these proteins might be related to cold stress.
【作者单位】：黑龙江八一农垦大学动物科技学院;
【基金】：“十三五”国家重点研发计划(2016YFD0501210) 国家自然科学基金面上项目(31272524) 农业部948计划重点项目(2011-G35)
【分类号】：S831

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